Preventing diseases in infants delivered via caesarean section

ABSTRACT

The present invention provides the use of a composition comprising non-digestible oligosaccharide for the manufacture of a composition for enteral administration to an infant delivered via caesarean section.

FIELD OF THE INVENTION

The present invention relates to methods for treatment and/or preventingdisorders in infants delivered via caesarean section.

BACKGROUND OF THE INVENTION

Human milk contains non-digestible oligosaccharides which specificallystimulate the growth of lactic acid producing bacteria, such as speciesbelonging to the genus Bifidobacterium and Lactobacillus and prevent thegrowth and/or adhesion to the intestinal wall of other (pathogenic)bacteria. Hence, when an infant receives human milk, the infant'sintestinal flora develops into a healthy flora rich in lactic acidproducing bacteria. The presence of a healthy intestinal flora improvesgut barrier maturation and/or gut barrier integrity, stimulates theformulation of mucus, inhibits pathogenic bacteria and stimulates theimmune system.

WO0008984 relates to a mixture of non-digestible carbohydrates forstimulating the health and enhancement of the healthy micro-organismspresent in the natural flora of the large intestine.

Before birth the intestinal tract of the infant is normally sterile.During vaginal delivery the intestinal tract of the infant is inoculatedwith vaginal and/or fecal bacteria of the mother, resulting in acolonization of the infant's gastrointestinal tract by bacteriaoriginating from the infant's environment.

Natren® produces the probiotic product Life Start® which is designedspecifically for infants and suitable for infants delivered viacaesarean section. Life Start® is made with Bifidobacterium infantis.Because the Life Start® product contains only one single Bifidobacteriaspecies, the benefits for the infant will be very limited.

SUMMARY OF THE INVENTION

The present inventors have found that infants delivered via caesareansection have an intestinal flora which is different from the intestinalflora of infants born via the vaginal route. Particularly, infants bornvia caesarean section have a reduced rate of intestinal colonization byBifidobacteria and have a less diverse Bifidobacterium intestinal floraregarding species than infants born via the vaginal route, particularlymissing Bifidobacterium breve, Bifidobacterium longum, Bifidobacteriuminfantis and Bifidobacterium bifidum.

It was further found that the intestinal flora of infants delivered viacaesarean section have a lower content of Bifidobacteria compared to theintestinal flora of infants delivered vaginally. Additionally it wasfound that the intestinal flora of infants born via caesarean sectionhas a high content of (undesirable) Escherichia coli 6 weeks afterdelivery.

The present inventors have found that these deficiencies in theintestinal flora of infants can be overcome by the administration ofnon-digestible oligosaccharides to the infant born via caesareansection, preferably included in the nutrition of the infant born viacaesarean section. Particularly, it was found that the administration ofnon-digestible oligosaccharides resulted in an increased content ofbifidobacteriam in infants delivered via caesarean section, and alsoreduces E. coli content. Overcoming such deficiencies results inimproved health and prevents and/or treats a variety of disorders.

A healthy and fully developed gastrointestinal flora has importantphysiological effects. One important aspect is that it reduces theoccurrence of (gastrointestinal) infections. Because infants deliveredvia a caesarean section lack a healthy flora, preventing infection isparticularly important for these infants. These infants are normallydelivered in a hospital environment, which is a risk for pathogenicinfection due to the occurrence of nosocomial bacteria. Additionally,the impaired development of a healthy intestinal flora results in fastercolonization of pathogenic bacteria compared to a situation where theinfants intestinal tract is inoculated by maternal bacteria.

The present invention particularly aims to decrease the number andseverity of (gastrointestinal) infections in infants born via caesareansection, by (i) stimulating the growth of beneficial bacteria,preferably lactic acid producing bacteria, (ii) decreasing the growth ofpathogenic bacteria; and/or (iii) decreasing the adhesion of pathogenicbacteria to the intestinal epithelial cells and/or intestinal mucus.

The present inventors have found that (gastrointestinal) infections canbe prevented and treated by non-digestible oligosaccharides,particularly galacto-oligosaccharides. The present non-digestibleoligosaccharides are suitable for enteral (particularly oral)administration, making it easy to include these in infant milk formula.The present non-digestible oligosaccharide reduces the occurrence of theinfections, as well as reduces the severity of infections due toreduction of intestinal concentrations of pathogens particularlyStaphylococcus aureus, Staphylococcus epidermidis, Streptococcushaemolyticus, Streptococcus, Clostridium difficile, Bacillus subtilis,Pseudomonas aeruginosa, Enterobacter, Klebsiella, Acinetobacter,Proteus, Aeromonas, and Escherichia coli.

The occurrence and severity of infection in infants delivered viacaesarean section can be even further reduced by providing a mixture ofat least two non-digestible oligosaccharides differing in structureand/or degree of polymerization (DP), and even further by providing inaddition to a non-digestible neutral oligosaccharide an acidicoligosaccharide, particularly uronic acid oligosaccharide. A mixture ofnon-digestible oligosaccharides differing in structure and/or DPsynergistically stimulates the development of a healthy gastrointestinalflora. Galacto-oligosaccharides, and/or fructo-oligosaccharides areparticularly suitable as these were found to stimulate the growth of B.longum, B. breve and/or B. infantis (Example 2). Uronic acidoligosaccharides prevent adhesion of pathogenic bacteria to theintestinal wall, thereby further preventing and/or treating infectionscaused by pathogens.

In addition to a reduced occurrence of infection, other disorders areprevented and/or treated in the infants delivered via caesarean sectionby the present method. Particularly disorders such as allergy and eczemaare prevented and/or treated by stimulating a healthy intestinal flora.

In a further aspect the present invention can be suitably brought topractice by incorporation of the present active ingredients in anutritional composition. Such composition can be administered to theinfant without posing a heavy burden on the infant delivered viacaesarean section.

DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

The present method provides the use of a composition comprisingnon-digestible oligosaccharides for the manufacture of a composition for(i) treatment and/or prevention of disorder in infants delivered viacaesarean section and/or (ii) the stimulation of health in infantsdelivered via caesarean section

In a further aspect the present invention provides a method for (i)treatment and/or prevention of disorders in infants delivered viacaesarean section and/or (ii) the stimulation of health in infantsdelivered via caesarean section, said method comprising administeringnon-digestible oligosaccharide to an infant delivered via caesareansection, preferably a method for the prevention and/or treatment ofinfection; and/or the prevention and/or treatment of allergy in infantsdelivered via caesarean section.

In a further aspect the present invention provides a method forstimulating the development of a healthy intestinal flora and/ordecreasing the occurrence of intestinal pathogens in an infant born viacaesarean section comprising the steps of: a) admixing I) anutritionally or pharmaceutically acceptable liquid; and II) a drycomposition, wherein the dry composition II comprises non-digestibleoligosaccharide and b) administering the composition obtained in step a)to the infant.

In still a further aspect the present invention provides a method forstimulating the development of a healthy intestinal flora and/ordecreasing intestinal pathogens in an infant delivered via caesareansection, comprising administering to the infant a composition comprisinga non-digestible oligosaccharide.

In a further aspect the present invention provides a method forproviding nutrition to an infant delivered via caesarean section, saidmethod comprising the steps of: a) admixing a composition comprisingnon-digestible oligosaccharide with a nutrition to be administered tothe infant delivered via caesarean section; and b) administering themixture obtained in step a) to the infant delivered via caesareansection. The invention also relates to a composition comprising at least5 wt. % non-digestible oligosaccharide based on dry weight of thecomposition and at least 1 wt. % uronic acid oligosaccharide based ondry weight of the composition.

Caesarean Section

The present invention relates to the enteral administration of acomposition comprising non-digestible oligosaccharides to infantsdelivered via caesarean section. A caesarean section (c-section) is asurgical procedure where an infant is delivered through an incision madein the mother's abdominal wall, and then through the wall of the uterus.A caesarean section is usually performed when it is safer for the motheror the infant than a vaginal delivery. Alternatively, a woman may chooseto have a caesarean section rather than deliver her infant vaginally.

Non-Digestible Oligosaccharides

The present composition comprises a non-ingestible oligosaccharide,which preferably stimulates the growth of the intestinal lactic acidproducing bacteria, particularly Bifidobacteria and/or the Lactobacilli.

The term “oligosaccharide” as used in the present invention refers tosaccharides with a degree of polymerization (DP) of 2 to 250, preferablya DP 2 to 100, more preferably 2 to 60, even more preferably 2 to 10. Ifthe oligosaccharide with a DP of 2 to 100 is included in the presentcomposition, this includes compositions which contain oligosaccharideswith a DP between 2 and 5, a DP between 50 and 70 and a DP of 7 to 60.The term “non-digestible oligosaccharide” as used in the presentinvention refers to oligosaccharides which are not or only partiallydigested in the intestine by the action of acids or digestive enzymespresent in the human upper digestive tract (small intestine and stomach)but which are fermented by the human intestinal flora. For example,sucrose, lactose, maltose and maltodextrins are considered digestible.Preferably the non-digestible oligosaccharide is a non-digestibleneutral oligosaccharide. The term “neutral oligosaccharide” as used inthe present invention refers to oligosaccharides wherein more than 75%of the saccharides units are selected from the group consisting ofglucose, fructose, galactose, mannose, ribose, rhamnose, arabinose, andxylose, preferably more than 85%, more preferably more than 95%, evenmore preferably more than 99%. Preferred neutral oligosaccharides aretransgalacto-oligosaccharides and fructo-oligosaccharides.

Preferably the present non-digestible oligosaccharide is a prebioticoligosaccharide. The term “prebiotic oligosaccharide” refers to anon-digestible oligosaccharide that beneficially affects the host byselectively stimulating the growth and/or activity of one or a limitednumber of probiotic bacterial species in the colon.

Preferably the present non-digestible oligosaccharide is soluble. Theterm “soluble” as used herein, when having reference to apolysaccharide, fiber or oligosaccharide, means that the substance is atleast soluble according to the method described by L. Prosky et al., J.Assoc. Off. Anal. Chem. 71, 1017-1023 (1988).

Preferably the present composition comprises at least one non-digestibleoligosaccharide selected from the group consisting ofgalacto-oligosaccharides, non-digestible dextrins,xylo-oligosaccharides, arabino-oligosaccharides, gluco-oligosaccharides(including gentio-oligosaccharides and cyclodextrins),chito-oligosaccharides, fuco-oligosaccharides, manno-oligosaccharides,isomalto-oligosaccharides fructo-oligosaccharides (including inulin),galactomanno-oligosaccharides, glucomanno-oligosaccharides, andarabinogalacto-oligosaccharides; more preferably at leastgalacto-oligosaccharides and/or fructo-oligosaccharides, most preferablyat least galacto-oligosaccharides.

The term “fructo-oligosaccharide” as used herein refers to anon-digestible polysaccharide carbohydrate comprising a chain of atleast 2 β-linked fructose units, with a DP of 2 to 250, preferably 7 to100, more preferably 20 to 60. Preferably inulin is used. Inulin isavailable under the tradename “Raftilin HP®”, (Orafti). The average DPof the present fructo-oligosaccharide is preferably at least 7, morepreferably at least 10, preferably below 100. The fructo-oligosaccharideused preferably has the (majority of) fructose units linked with aβ(2→1) linkage. Other terms for fructooligosaccharides include inulin,fructopolysaccharide, polyfructose, fructans and oligofructose. Thepresent composition preferably comprises fructo-oligosaccharides with aDP of 2 to 100.

Non-digestible dextrins refer to digestion-resistant (malto)dextrins ordigestion-resistant polydextrose which preferably have a DP of 10 to 50,preferably between 10 and 20. The non-digestible dextrins preferablycomprise α(1→4), α(1→6) glucosidic bonds and 1→2 and 1→3 linkagesNon-digestible dextrins are for example available under the tradename“Fibersol 2®” from Matsutami Industries or Litesse® from Danisco.

The present inventors found that galacto-oligosaccharides can beadvantageously used in the present composition, because theseoligosaccharides where particularly effective in stimulating the growthof Bifidobacteria. Hence, in a preferred embodiment the presentcomposition comprises galacto-oligosaccharides. The term“galacto-oligosaccharide” as used herein refers to a non-digestibleoligosaccharide, wherein at least 30% of the saccharide units aregalactose units, preferably at least 50%, more preferably at least 60%.The present composition preferably comprises galacto-oligosaccharideswith a DP of 2-100, more preferably a DP of 2-10. Preferably thesaccharides of the galacto-oligosaccharide are β-linked, as is the casein human milk oligosaccharides.

Preferably the present composition comprises a galacto-oligosaccharideselected from the group consisting of transgalacto-oligosaccharides,lacto-N-tetraose (LNT) and lacto-N-neotetraose (neo-LNT). In aparticularly preferred embodiment the present method comprises theadministration of transgalacto-oligosaccharide ([galactose]_(n)-glucose;wherein n is an integer between 1 and 60, i.e. 2, 3, 4, 5, 6, . . . ,59, 60; preferably n is 2, 3, 4, 5, 6, 7, 8, 9 and/or 10).Transgalacto-oligosaccharides (TOS) are for example sold under thetrademark Vivinal™ (Borculo Domo Ingredients, Netherlands).

The present composition preferably comprises 0.5 to 75 grams of thenon-digestible oligosaccharides per 100 gram dry weight, preferablybetween 0.5 and 50 grams. The present composition preferably comprises0.1 to 75 grams of the galacto-oligosaccharides per 100 gram dry weight,preferably between 0.1 and 50 grams.

The present method preferably comprises the administration of a servingcomprising between 0.05 and 25 grams non-digestible oligosaccharide,preferably between 0.1 and 5 grams. The present method preferablycomprises the administration of a serving comprising between 0.05 and 25grams galacto-oligosaccharides, preferably between 0.1 and 5 gramgalacto-oligosaccharides.

The present inventors have also found that a mixture of a long chainnon-digestible (neutral) oligosaccharides and short chain non-digestible(neutral) oligosaccharides synergistically stimulate the growth of ahealthy intestinal flora, particularly Bifidobacteria and reduces theoccurrence of E. coli in infants delivered via caesarean section.

The present composition thus preferably comprises at least twonon-digestible (neutral) oligosaccharides with different average degreesof polymerization (DP). Preferably the weight ratios:

-   a. (non-digestible (neutral) oligosaccharides with DP 2 to 5):    (non-digestible (neutral) oligosaccharides with DP 6, 7, 8, and/or    9)>1; and/or-   b. (non-digestible (neutral) oligosaccharides with DP 10 to 60):    (non-digestible (neutral) oligosaccharides with DP 6, 7, 8, and/or    9)>1

Preferably both weight ratios are above 2, even more preferably above 5.

For further improvement, the present non-digestible oligosaccharidepreferably has a relatively high content of short chainoligosaccharides, as these strongly stimulate the growth ofBifidobacteria. Hence, preferably at least 10 wt. % of thenon-digestible oligosaccharides in the present composition has a DP of 2to 5 (i.e. 2, 3, 4, and/or 5) and at least 5 wt. % has a DP of 10 to 60.Preferably at least 50 wt. %, more preferably at least 75 wt. % of thenon-digestible neutral oligosaccharides have a DP of 2 to 9 (i.e. 2, 3,4, 5, 6, 7, 8, and/or 9).

To improve the biodiversity and stimulate the growth of multipleintestinal organisms, the present composition preferably comprises twonon-digestible oligosaccharides with a different structure. The presentcomposition preferably comprises at least two different non-digestible(neutral) oligosaccharides, wherein the oligosaccharides have a homologyin saccharide units below about 90%, preferably below 50%, even morepreferably below 25%, even more preferably below 5%. The term “homology”as used in the present invention is the cumulative of the percentage ofsame saccharide unit in the different oligosaccharides. For example,oligosaccharide 1 (OL1) has the structure fruc-fruc-glu-gal, and thuscomprises 50% fruc (fructose), 25% gal (galactose) and 25% glu(glucose). Oligosaccharide 2 (OL2) has the structure fruc-fruc-glu, andthus comprises 66% fruc, 33% glu. The different oligosaccharides thushave a homology of 75% (50% fruc+25% glu).

The present composition preferably comprises galacto-oligosaccharidesand fructo-oligosaccharides, more preferablytransgalacto-oligosaccharides with a DP of 2-7 andfructo-oligosaccharides with a DP of 10-100.

Uronic Acid Oligosaccharides

The present composition preferably comprises uronic acidoligosaccharide, more preferably a combination of non-digestible neutraloligosaccharide and uronic acid oligosaccharide. The uronic acidoligosaccharide (further) reduces the adherence of pathogens to theintestinal epithelia and suppresses colonization by intestinalpathogens.

The term uronic acid oligosaccharide as used in the present inventionrefers to an oligosaccharide wherein preferably at least 25%, preferablyat least 50% of the monosaccharide units present in the oligosaccharideis one selected from the group consisting of guluronic acid, mannuronicacid, iduronic acid, riburonic acid, galacturonic acid and glucuronicacid. In a preferred embodiment the ironic acid oligosaccharidecomprises at least 50% galacturonic acid based on total uronic acidunits in the uronic acid oligosaccharide. The uronic acidoligosaccharides used in the invention are preferably prepared frompectin, pectate, alginate, chondroitine, hyaluronic acids, heparine,heparane, bacterial carbohydrates, sialoglycans, fucoidan,fuco-oligosaccharides and/or carrageenan, more preferably from pectinand/or alginate, even more preferably from pectin, most preferablypolygalacturonic acid. The present uronic acid oligosaccharide ispreferably a pectin degradation product and/or alginate degradationproduct. Preferably the pectin degradation product is a pectinhydrolysate (prepared by hydrolysis) and/or pectin lysate (prepared bybeta-elimination). The pectin degradation product is preferably preparedfrom fruit and/or vegetable pectin, more preferably apple pectin, citruspectin and/or sugar beet pectin, more preferably from apple, citrusand/or sugar beet pectin. The pectin degradation product is preferablyprepared with lyases and/or variations of the temperature and pressure,more preferably with pectin lysate, i.e. by beta-elimination. The pectindegradation product is preferably a pectin lysate

Preferably the present composition comprises uronic acid oligosaccharidewith a DP of 2 to 250, more preferably a DP of 2 to 100, even morepreferably a DP of 2 to 50, most preferably a DP of 2 to 20. Preferablythe present composition comprises between 25 and 100 wt. %, morepreferably between 50 and 100 wt. % uronic acid oligosaccharide with aDP of 2 to 250 based on total weight of uronic acid in the composition,more preferably a DP of 2 to 100, even more preferably a DP of 2 to 50,most preferably a DP of 2 to 20.

The present uronic acid oligosaccharide is preferably obtainable byenzymatic digestion of pectin with pectin lyases, pectic lyase,endopolygalacturonase and/or pectinase.

In a preferred embodiment at least one of the terminal hexuronic acidunits of the uronic acid oligosaccharide has a double bond, which ispreferably situated between the C₄ and C₅ position of the terminalhexuronic acid unit. The double bond effectively protects againstattachment of the pathogenic bacteria to the intestinal epithelialcells. Preferably one of the terminal hexuronic acid units comprises thedouble bond. The double bond at terminal hexuronic acid unit can forexample be obtained by enzymatic hydrolysis of pectin with lyase.

In a further embodiment, a mixture of uronic acid oligosaccharides isused, which have a different DP and/or comprise both unsaturated andsaturated terminal hexuronic acid units. Preferably at least 5%, morepreferably at least 10%, even more preferably at least 25% of theterminal hexuronic acid units of the uronic acid oligosaccharide is anunsaturated hexuronic acid unit. As each individual uronic acidoligosaccharide preferably comprises only one unsaturated terminalhexuronic acid unit, preferably less than 50% of the terminal hexuronicacid units is an unsaturated hexuronic acid unit (i.e. comprises adouble bond). A mixture of uronic acid oligosaccharides preferablycontains between 2 and 50% unsaturated terminal hexuronic acid unitsbased on the total amount of terminal hexuronic acid units, preferablybetween 10 and 40%.

The uronic acid oligosaccharide can be derivatized. The uronic acidoligosaccharide may be methoxylated and/or amidated. In one preferredembodiment the uronic acid oligosaccharides are characterized by adegree of methoxylation above 20%, preferably above 30% even morepreferably above 70%. As used herein, “degree of methoxylation” (alsoreferred to as DE or “degree of esterification”) is intended to mean theextent to which free carboxylic acid groups contained in the uronic acidoligosaccharide have been esterified (e.g. by methylation). In anotherpreferred embodiment the uronic acid oligosaccharides have a degree ofmethylation above 20%, preferably above 30%, even more preferably above70%.

The present composition preferably comprises between 0.01 and 10 gramsuronic acid oligosaccharide with a DP of 2 to 250 per 100 gram dryweight of the present composition, more preferably between 005 and 6gram, even more preferably 0.2 to 2 gram per 100 gram dry weight. Thepresent composition preferably comprises between 0.01 and 10 gramsgalacturonic acid oligosaccharide with a DP of 2 to 250 (more preferablya DP of 2 to 100) per 100 gram dry weight of the present composition,more preferably between 0.05 and 6 grain, even more preferably 0.2 to 2gram.

The uronic acid oligosaccharides are preferably non-digestible in theupper human intestinal tract and water-soluble (according to the methoddisclosed in L. Prosky et al, J. Assoc. Anal. Chem. 71: 1017-1023,1988). The uronic acid oligosaccharides are preferably fermentable bythe intestinal flora. The uronic acid oligosaccharides of the inventionadvantageously reduce the adhesion of pathogenic micro-organisms to theintestinal epithelial cells, thereby reducing colonization of(nosocomial) pathogenic bacteria in the colon of the infant delivered bycaesarean section. Furthermore, the uronic acid oligosaccharides of thepresent invention preferably stimulate the formation of a healthyintestinal flora and are fermented, resulting in a production ofintestinal organic acids and a reduction of intestinal pH, which inhibitthe growth of (nosocomial) pathogenic bacteria.

Lactic Acid Producing Bacteria

The present composition preferably comprises lactic acid producingbacteria, either living or dead. Lactic acid producing bacteria arepreferably provided as a mono- or mixed culture of live microorganisms.The present composition preferably comprises 10² to 10¹³ colony formingunits (cfu) of lactic acid producing bacteria per gram dry weight of thepresent composition, preferably 10² to 10¹², more preferably 10⁵ to10¹⁰, most preferably from 10⁴ to 5×10⁹ cfu.

Preferably the present composition comprises bacteria of the genusLactobacillus or Bifidobacterium. Preferably the composition comprisesat least one Bifidobacterium selected from the group consisting of B.longum, B. breve, B. infantis, B. catenulatum, B. pseudocatenulatum, B.adolescentis, B. animalis, B. gallicum, B. lactis and B. bifidum, morepreferably B. breve, B. infantis, B. bifidum, B. catenulatum, B. longum,more preferably B. longum and B. breve, most preferably B. breve.Preferably the composition comprises at least two differentBifidobacterium species, subspecies or strains. The present compositionpreferably comprises at least one, more preferably at least two, evenmore preferably at least three, most preferably at least four differentBifidobacterium species. The present composition preferably comprises atleast one, more preferably at least two, even more preferably at leastthree, most preferably at least four different Bifidobacterium strains.Preferably the present composition comprises at least B. longum and B.breve. The above-mentioned combinations commonly aim to increase thediversity and/or the quantity of microorganisms in the intestine of thecaesarean section delivered infant. This beneficially affects theinfant, proving numerous health benefits.

Preferably the present composition comprises a Lactobacillus selectedfrom the group consisting of L. casei, L. reuteri, L. paracasei, L.rhamnosus, L. acidophilus, L. johnsonii, L. lactis, L. salivarius, L.crispatus, L. gasseri, L. zeae, L. fermentum and L. plantarum, morepreferably L. casei, L. paracasei, L. rhamnosus, L. johnsonii, L.acidophilus, L. fermentum and most preferably L. paracasei. Even morepreferably the present composition comprises Bifidobacterium breveand/or Lactobacillus paracasei, because the growth of these bacteria inimpaired in the intestine of formula fed infants compared to breast fedinfants. The further increased biodiversity will have a stimulatoryeffect on health of the newborn delivered by caesarean section.

Long-Chain Polyunsaturated Fatty Acids

The present composition preferably comprises long chain polyunsaturatedfatty acids (LC-PUFA). LC-PUFA are fatty acids or fatty acyl chains witha length of 20 to 24 carbon atoms, preferably 20 or 22 carbon atomscomprising two or more unsaturated bonds. More preferably the presentcomposition comprises eicosapentaenoic acid (EPA, n-3), docosahexanoicacid (DHA, n-3) and/or arachidonic acid (ARA, n-6). These LC-PUFAeffectively reduce intestinal tight junction permeability. Reduced tightjunction permeability reduced the occurrence of infection and/or reducespassage of allergens. Hence incorporation of EPA, DHA and/or ARA in thepresent composition improves intestinal barrier integrity, which is ofutmost important for babies delivered via a caesarean section sincethese babies have a less developed intestinal flora and hence a slowermaturing gut barrier. The incorporation of these LC-PUFA's will furthercontribute (synergistically) to the reduced occurrence and severity ofinfections and allergy.

Since low concentration of ARA, DHA and/or EPA are already effective inreducing tight junction permeability, the content of LC-PUFA with 20 and22 carbon atoms in the present composition preferably does not exceed 15wt. % of the total fat content, preferably does not exceed 10 wt. %,even more preferably does not exceed 5 wt. % of the total fat content.Preferably the present composition comprises at least 0.1 wt. %,preferably at least 0.25 wt. %, more preferably at least 0.6 wt. %, evenmore preferably at least 0.75 wt. % LC-PUFA with 20 and 22 carbon atomsbased on total fat content. For the same reason, the EPA contentpreferably does not exceed 5 wt. % of the total fat, more preferablydoes not exceed 1 wt. %, but is preferably at least 0.03 wt. %, morepreferably at least 0.05 wt. % of the total fat. The DHA contentpreferably does not exceed 5 wt. %, more preferably does not exceed 1wt. %, but is at least 0.1 wt. % of the total it. As ARA was found to beparticularly effective in reducing tight junction permeability, thepresent composition comprises relatively high amounts, preferably atleast 0.1 wt. %, even more preferably at least 0.25 wt. %, mostpreferably at least 0.35 wt. % of the total fat. The ARA contentpreferably does not exceed 5 wt. %, more preferably does not exceed 1wt. % of the total fat. When the present enteral composition comprisesEPA and DHA are advantageously added to balance the action of ARA, e.g.reduce the potential pro-inflammatory action of ARA metabolites. Excessmetabolites from ARA may cause inflammation. Hence, the presentcomposition preferably comprises ARA, EPA and DHA, wherein the weightratio ARA/DHA preferably is above 0.25, preferably above 0.5, even morepreferably above 1. The ratio is preferably below 25, more preferablybelow 15. The weight ratio ARA/EPA is preferably between 1 and 100, morepreferably between 5 and 20.

The present composition preferably comprises between 5 and 75 wt. %polyunsaturated fatty acids based on total fat, preferably between 10and 50 wt. %.

The content of LC-PUFA, particularly the LC-PUFA with 20 and 22 carbonatoms, preferably does not exceed 3 wt. % of the total fat content as itis desirable to mimic human milk as closely as possible. The LC-PUFA maybe provided as free fatty acids, in triglyceride form, in diglycerideform, in monoglyceride form, in phospholipid form, or as a mixture ofone of more of the above. The present composition preferably comprisesat least one of ARA and DHA in phospholipid form.

Nucleotides

Preferably the present composition comprises nucleotides and/ornucleosides, more preferably nucleotides. Preferably, the compositioncomprises cytidine 5′-monophosphate, uridine 5′-monophosphate, adenosine5′-monophosphate, guanosine 5′-1-monophosphate, and/or inosine5′-monophosphate, more preferably cytidine 5′-monophosphate, uridine5′-monophosphate, adenosine 5′-monophosphate, guanosine5′-monophosphate, and inosine 5′-monophosphate.

Preferably the composition comprises 5 to 100, more preferably 5 to 50mg, most preferably 10 to 50 mg nucleotides and/or nucleosides per 100gram dry weight of the composition. The presence of nucleotides and/ornucleotides advantageously enhances gut growth and maturation in theinfant, which is of crucial importance in infants delivered by caesareansection. The nucleotides and/or nucleosides further stimulate the immunesystem thereby enhancing protection against a high load of intestinalpathogens such as E. coli. The nucleotides and/or nucleosides are deemedto act synergistically with the other ingredients of the presentcomposition.

Formulae

The present composition is preferably enterally administered, morepreferably orally.

The present composition is preferably a nutritional formula, preferablyan infant formula. The present composition can be advantageously appliedas a complete nutrition for infants. The present composition preferablycomprises a lipid component, protein component and carbohydratecomponent and is preferably administered in liquid form. The presentinvention includes dry food (e.g. powders) which are accompanied withinstructions as to admix said dry food mixture with a suitable liquid(e.g. water).

The present invention advantageously provides to a composition whereinthe lipid component provides 5 to 50% of the total calories, the proteincomponent provides 5 to 50% of the total calories, and the carbohydratecomponent provides 15 to 90% of the total calories. Preferably, in thepresent composition the lipid component provides 35 to 50% of the totalcalories, the protein component provides 7.5 to 12.5% of the totalcalories, and the carbohydrate component provides 40 to 55% of the totalcalories. For calculation of the % of total calories for the proteincomponent, the total of energy provided by the proteins, peptides andamino acids needs to be taken into account.

The present composition preferably comprises at least one lipid selectedfrom the group consisting of animal lipid (excluding human lipids) andvegetable lipids. Preferably the present composition comprises acombination of vegetable lipids and at least one oil selected from thegroup consisting of fish oil, animal oil, algae oil, fungal oil, andbacterial oil. The present composition comprising non-digestibleoligosaccharides excludes human milk.

The protein component used in the nutritional preparation are preferablyselected from the group consisting of non-human animal proteins(preferably milk proteins), vegetable proteins (preferably soy proteinand/or rice protein), free amino acids and mixtures thereof. Cow's milkderived nitrogen source. The present composition preferably containscasein, whey, hydrolysed casein and/or hydrolysed whey protein.Preferably the protein comprises intact proteins, more preferably intactbovine whey proteins and/or intact bovine casein proteins. As thepresent composition is suitably used to reduce the allergic reaction inan infant, the protein of is preferably selected from the groupconsisting of hydrolyzed milk protein. Preferably the presentcomposition comprises hydrolyzed casein and/or hydrolyzed whey protein,vegetable protein and/or amino acids. The use of these proteins furtherreduced the allergic reactions of the infant. The use of thesehydrolysed proteins advantageously improves the absorption of thedietary protein component by the immature intestine of the infantdelivered by caesarean section.

The present composition preferably contains digestible carbohydratesselected from the group consisting of sucrose, lactose, glucose,fructose, corn syrup solids, starch and maltodextrins, more preferablylactose.

Stool irregularities (e.g. hard stools, insufficient stool volume,diarrhea) is an important problem in babies delivered via caesareansection. This may be caused by the high content of E. coli in the feces.It was found that stool problems may be reduced by administering thepresent non-digestible oligosaccharides in liquid food with anosmolality between 50 and 500 mOsm/kg, more preferably between 100 and400 mOsm/kg. The reduced stool irregularities enhance the colonizationand development of a healthy intestinal flora.

In view of the above, it is also important that the liquid food does nothave an excessive caloric density, however still provides sufficientcalories to feed the subject. Hence, the liquid food preferably has acaloric density between 0.1 and 2.5 kcal/mL, even more preferably acaloric density of between 0.5 and 1.5 kcal/ml, most preferably between0.6 and 0.8 kcal/ml.

Concentrated Non-Digestible Oligosaccharides

According to a further preferred embodiment, the present inventionsprovides a method for providing nutrition to an infant delivered viacaesarean section, wherein a composition containing oligosaccharides,preferably a concentrated compositions containing high amounts ofoligosaccharides, is admixed to the nutrition to be administered to theinfant born via caesarean section. Providing a concentrated form enablesthe addition of the present non-digestible oligosaccharides to bothsynthetic infant milk formula but also breast milk.

The present invention thus provides a method for providing nutrition toan infant delivered via caesarean section, said method comprising thesteps of:

-   a) admixing a composition comprising non-digestible oligosaccharide    with a nutrition to be administered to the infant delivered via    caesarean section; and-   b) administering the mixture obtained in step a) to the infant    delivered via caesarean section.

Preferably the nutrition as referred to in step a) of the method isinfant milk formula or human breast milk. The composition comprisingnon-digestible oligosaccharides as used in step a) is a concentratedcomposition of non-digestible oligosaccharides or in other words is acomposition comprising non-digestible oligosaccharides in high amounts.

The composition comprising non-digestible oligosaccharide as used instep a) in the above-described method, preferably comprises at least 5wt. %, preferably at least 10 wt. %, more preferably at least 25 wt. %non-digestible oligosaccharide based on dry weight of the composition,wherein the non-digestible oligosaccharide is preferably selected fromthe group consisting of galacto-oligosaccharides, non-digestibledextrins, xylo-oligosaccharides, arabino-oligosaccharides,gluco-oligosaccharides (including gentio-oligosaccharides andcyclodextrins), chito-oligosaccharides, fuco-oligosaccharides,manno-oligosaccharides, isomalto-oligosaccharide andfructo-oligosaccharide (including inulins). This composition can beimproved by combining it with one or more of the components as disclosedabove, preferably one or more or all selected from the group of uronicacid oligosaccharides, LC-PUFA, nucleotides and probiotic bacteria.

The present invention also provides a composition which can be suitablyused in the present method for admixing to a nutrition, i.e. acomposition which comprises a high concentrations of non-digestibleneutral oligosaccharide. The composition comprises at least 5 wt. %(preferably at least 10 wt. %) non-digestible oligosaccharide based ondry weight of the composition, wherein the non-digestibleoligosaccharide is preferably selected from the group consisting ofgalacto-oligosaccharides, non-digestible dextrins,xylo-oligosaccharides, arabino-oligosaccharides, gluco-oligosaccharides(including gentio-oligosaccharides and cyclodextrins),chito-oligosaccharides, fuco-oligosaccharides, manno-oligosaccharides,isomalto-oligosaccharide and fructo-oligosaccharide (including inulins);and at least 1 wt. % (preferably at least 10 wt. %) of an uronic acidoligosaccharide, preferably as defined above, based on dry weight of thecomposition. The composition preferably contain galactooligosaccharidesand/or fructooligosaccharides.

The composition is preferably designed to be added to a single servingof infant nutrition. The cumulative weight of the non-digestibleoligosaccharide in the present composition is preferably between 0.1 and10 gram, preferably between 0.2 and 5 grain per serving. The compositionis preferably packed per serving, i.e. in a unit dose of one serving,preferably in the form of a sachet. On serving of the compositionpreferably has a dry weight of 0.5 to 25 grams, preferably between 1 and10 grains.

Application

The present invention provides (i) the treatment and/or prevention of adisorder in infants delivered via caesarean section and/or (ii) thestimulation of health in infants delivered via caesarean section. Thedisorder is preferably selected from the group consisting of intestinaldisorders caused by low bifidogenic flora. Preferably the disorder isselected from the group of infection and allergy. The present inventionpreferably provides a method for the prevention and/or treatment ofinfections and/or infection disorders, particularly gastrointestinalinfections, more preferably the treatment and/or prevention ofinfections caused by one or more micro-organisms selected from the groupconsisting of Staphylococcus (especially S. aureus, S. epidermidis, S.haemolyticus), Streptococcus (especially Streptococcus group B),Clostridium (especially C. difficile), Bacillus (especially B. subtilis,Pseudomonas (especially P. aeruginosa), Enterobacter, Klebsiella,Acinetobacter, Proteus, Aeromonas, and Escherichia coli, preferablyEscherichia coli (E. coli). Preferably, the present composition is usedin a method for treatment and/or prevention of intestinal infection,urinary tract infection, intestinal inflammation and/or diarrhea ininfants delivered by caesarean section. Preferably the presentcomposition is used in a method for modulating the immune system ininfants born via caesarean section. Preferably the present compositionis used in a method modulate the immune system, preferably resulting inthe protection of the infant from allergy.

The present composition is preferably administered to the infantdelivered via caesarean section in the first year of life, preferablywithin 3 months after birth, even more preferably within two weeks afterbirth, even more preferably within 100 hours, more preferably within 72hours, most preferably within 48 hours after birth.

The present invention also provides a method for stimulating thedevelopment of a healthy intestinal flora in an infant comprising stepA: admixing I) a nutritionally or pharmaceutically acceptable liquid;and II) a dry composition, wherein the dry composition II comprisesnon-digestible oligosaccharides; and step B) administering thecomposition obtained in step A to an infant born via caesarean section.

Administration of the present composition results in an improvedintestinal flora and/or in the formation of organic acids as metabolicend products of microbial fermentation. An increased amount of organicacids results in an increased mucus production, improves gut maturationand/or and increased gut barrier. Hence, in a further aspect, thepresent invention provides a method for treatment and/or prevention ofallergy particularly food allergy), atopic eczema (e.g. atopicdermatitis), asthma, allergic rhinitis, allergic conjunctivitis ininfants delivered by caesarean section, said method comprisingadministering to the infant a composition comprising the presentnon-digestible (neutral) oligosaccharides.

Furthermore, administration of the present composition strengthens theimmune system. In a further aspect, the present invention thereforeprovides a method for treatment and/or prevention of systemicinfections, otitis and/or respiratory infections in infants delivered bycaesarean section.

EXAMPLES Example 1 Molecular Characterization of Intestinal Microbiotain Infants Born by Vaginal Delivery Vs. Caesarean Delivery

In the present study the influence of mode of delivery (caesareandelivery versus vaginal delivery) on the intestinal microbialcomposition at the third day of life by was studied using by PCRamplification with species-specific primers for ten Bifidobacteriumspecies, three Ruminococcus species and one Bacteroides species.

The microbial DNA was extracted and analyzed according to Favier et al,Environ Microbiol 2002; 68:219-226 and Satokari et al, Appl EnvironMicrobiol 2001; 67:504-513; Satorkari et al System Appl Microbiol 2003;26:572-584.

The results of the Bifidobacterium and other species detected in faecalsamples of 21 newborns after caesarean delivery obtained at the 3rd dayof life are given in Table 1. Table 2 gives the Bifidobacterium andother species detected in faecal samples of 21 newborns after vaginaldelivery obtained at the 3rd day of life. No signal specific for thespecies B. dentium, B. angulatum, B. lactis, Ruminococcus bromii,Ruminococcus callidus and Ruminococcus obeum was observed in the faecesof infants delivered by caesarean section as well as of vaginallydelivered infants.

It can be concluded that the microbial flora of an infant born viacaesarean section differs from that of an infant born via the vaginalroute. Not only is the amount of bifidobacteria and other speciesquantitatively much lower, also on a species level the flora ofcaesarean section delivered infants is less diverse. SinceBifidobacterium is the dominant genus in infant's flora these resultscan also be generalised to less intestinal bacteria and a less diverseintestinal flora in caesarean section delivered infants, leaving theintestine more susceptible to colonisation by (nosocomial) pathogens.

These results are indicative for the advantageous use of the compositionand method according to the present invention, e.g. a method for feedingbabies born via caesarean section, decreasing intestinal pathogens,stimulating a healthy intestinal flora and consequently preventinginfection, stimulating a healthy immune system, and stimulating gutmaturation.

TABLE 1 Caesarean section delivered infants B. B. B. B. catenu- B. B. B.Bacteroides NEWBORN breve infantis bifidum latum group adolescentislongum gallicum fragilis 1 − − − − − − − − 2 − − − − − ++ − − 3 − − − −− − − − 4 − − − − − − − − 5 − − − − − − − − 6 − − − − − − − − 7 − − − −− − − − 8 − − − − − − − − 9 − − − − − − − − 10 − − − − − − ++ − 11 − − −− − − − − 12 − − − − − − − − 13 − − − − − − − − 16 − − − − − − − − 17 −− − − − − − − 18 − − − − − − − − 19 − − − − − − − − 20 − − − − − − − −21 − − − − − − − − 22 − − − − − − − − 23 − − − − − − − − (−) = noamplification; (+/−) = weak amplification; (+) = positive amplification;(++) = strong amplification

TABLE 2 Vaginally delivered infants B. B. B. B. catenu- B. B. B.Bacteroides NEWBORN breve infantis bifidum latum group adolescentislongum gallicum fragilis  1a − + − − − ++ − −  2a +/− − ++ ++ − ++ − − 3a − − − + − − − −  4a +/− − − ++ + + − −  5a +/− − ++ ++ − ++ ++ −  6a− − +/− ++ − ++ − −  7a − − +/− ++ ++ − − −  8a ++ ++ − + ++ − − −  9a −− − + ++ + − − 10a ++ − − + + − − − 11a ++ − ++ ++ − ++ − + 12a + + + +− ++ − + 13a +/− − − + − + − − 16a − − − ++ − + − − 17a +/− − + + − + −− 18a +/− − + + − + − − 19a + − − + − + − − 20a − − − + − + − − 21a − −− + ++ + − − 22a − + − ++ − + − − 23a + − ++ ++ − + − − (−) = noamplification; (+/−) = weak amplification; (+) = positive amplification;(++) = strong amplification

Example 2 Effect of Non-Digestible Oligosaccharides on the Flora inCaesarean Delivered Infants

Infants were administered infant formula supplemented with 0.8 g/100 mlgalacto-oligosaccharides (GOS) with an average DP between 2 and 7 andfructo-oligosaccharides with (FOS, Raftilin HP®) (GFSF-group) orstandard infant formula without non-digestible oligosaccharides(SF-group).

The bifidobacterial content in the feces was determined. The percentageof the genus Bifidobacterium as a percentage of total bacteria in thefirst week was 4.3% in caesarean delivered infants (n=44) versus 19.8%in vaginally delivered infants (n=28). At 6 weeks the percentagebifidobacteria was 12.3% in the SF-group of caesarean delivered infants(n=21) and 17.2% in the GFSF-group of the caesarean infants (n=13). Thefecal pH of the SF-group of caesarean delivered infants was 7.2 versus6.5 of the GFSF-group of caesarean delivered infants. The percentage E.coli was 11.8% in the SF group of caesarean delivered infants and 0% inthe GFSF-group of the caesarean delivered infants. (see Table 3)

These results indicate that administration of non-digestible neutraloligosaccharides to infants born via caesarean section (GFSF group)results in a more bifidogenic flora and a reduced content of potentiallypathogenic bacteria compared to infants born via caesarean section thatdo not receive non-digestible oligosaccharides (the SF group).Additionally the results indicate a reduced content of E. coli due tothe administration of non-digestible oligosaccharides. The results areindicative for the advantageous use of non-digestible oligosaccharides,particularly galacto-oligosaccharides and fructo-oligosaccharides in thepresent composition to further improve the present therapy in infantsborn via caesarean section.

TABLE 3 Percentage Bifidobacteria in vaginally and caesarean sectiondelivered infants fed a formula with (GFSF) or without (SF)non-digestible oligosaccharides. Bifidobacteria Bifidobacteria in firstweek after 6 weeks Fecal E. coli Infants (%) (%) pH (%) Vaginal delivery19.8 C-section SF 4.3 12.3 7.2 11.8 C-section GFSF 4.3 17.2 6.5 0

Example 3 Bifidogenic Effect of Non-Digestible Oligosaccharides

Infants were administered infant formula supplemented with 0.8 g/100 mlgalacto-oligosaccharides (GOS) with an average DP between 2 and 7 andfructo-oligosaccharides with (FOS, Raftilin HP®) (GFSF-group) orstandard infant formula without non-digestible oligosaccharides(SF-group).

The bifidobacterial content in the feces was determined. The percentageof the genus Bifidobacterium as a percentage of total bacteria at 6weeks was 47% in the SF and 68% in the GFSF group, respectively, whichdemonstrates that the GFSF group, fed a mixture of non-digestiblecarbohydrates, has a more bifidogenic flora compared to the SF group.Also on a species level the percentage Bifidobacterium infantis was 22%in the GFSF group and 18% in the SF group, B. longum was 3.7% in theGFSF group and 2.9% in the SF group and B. breve was 3.7% in the GFSFgroup and 2.3% in the SF group. The results are indicative for theadvantageous use of non-digestible neutral oligosaccharides,particularly galacto-oligosaccharides and fructo-oligosaccharides in thepresent composition to further improve the present therapy in infantsborn via caesarean section.

Example 4 Composition for Babies Born Via Caesarean Section

An infant formula composition comprising per 100 ml ready to feedformula: 1.6 g protein, 3.6 g fat, 6.4 g digestible carbohydrates(mainly lactose), 0.8 g non-digestible oligosaccharides of which 0.54 gtransgalacto-oligosaccharides, 0.06 g inulin, and 0.2 g pectinhydrolysate (prepared by lyase hydrolysis of citrus pectin). The packageand/or supporting material accompanying the product indicates that theproduct can be suitably used to a) prevent and/or treat infection ininfants delivered via caesarean section; b) prevent and/or treatinfection with E. coli in infants delivered via caesarean section; c)prevent and/or treat allergy in infants delivered via caesarean section;and/or d) modulate the immune system.

Example 5 Method for Feeding Babies Born Via Caesarean Section

Sachet which contains:

-   -   0.2 g uronic acid oligosaccharide (about 2 wt. % based on dry        weight);    -   2 g galacto-oligosaccharide and 0.3 g fructooligosaccharides        (about 20 wt. % based on dry weight);    -   Bifidobacterium breve and Bifidobacterium longum, about 5*10⁹        cfu of each strain;    -   Bulking agent

The invention claimed is:
 1. A method for treating and/or preventingatopic dermatitis in an infant delivered by caesarean section, themethod comprising feeding the infant a composition comprising: (a)intact and/or hydrolyzed protein and (b) 0.5 to 75 g of a non-digestibleneutral oligosaccharide per 100 g dry weight of the composition, whereinthe non-digestible neutral oligosaccharide consists ofgalacto-oligosaccharides, and, optionally, non-digestible dextrins,xylo-oligosaccharides, arabino-oligosaccharides, gluco-oligosaccharides,chito-oligosaccharides, fuco-oligosaccharides, manno-oligosaccharides,isomalto-oligosaccharides, fructo-oligosaccharides,arabinogalacto-oligosaccharides, glucomanno-oligosaccharides and/orgalactomanno-oligosaccharides, wherein (i) at least 50% of thesaccharide units of the galacto-oligosaccharide are galactose units,thereby treating and/or preventing atopic dermatitis.
 2. The methodaccording to claim 1, wherein the composition comprises: (i) a lipidcomponent that provides 5 to 50% of total calories of the composition;(ii) a protein component that provides 5 to 50% of total calories of thecomposition; and (iii) a carbohydrate component that provides 15 to 90%of the total calories of the composition.
 3. The method according toclaim 1, wherein the composition comprises long chain polyunsaturatedfatty acid and/or a nucleotide.
 4. The method according to claim 1,wherein the composition further comprises 10² to 10¹³ colony formingunits of lactic acid-producing bacteria per gram dry weight of thecomposition.
 5. The method according to claim 1, wherein the feeding ofsaid composition promotes the development of healthy intestinal floraand/or decreases the occurrence of intestinal pathogens in said infant.6. The method according to claim 1, wherein the composition is an infantformula.